Metabolic engineering of Pichia pastoris

Abstract

Besides its use for efficient production of recombinant proteins the methylotrophic yeast Pichia pastoris (syn. Komagataella spp.) has been increasingly employed as a platform to produce metabolites of varying origin. We summarize here the impressive methodological developments of the last years to model and analyze the metabolism of P. pastoris, and to engineer its genome and metabolic pathways. Efficient methods to insert, modify or delete genes via homologous recombination and CRISPR/Cas9, supported by modular cloning techniques, have been reported. An outstanding early example of metabolic engineering in P. pastoris was the humanization of protein glycosylation. More recently the cell metabolism was engineered also to enhance the productivity of heterologous proteins. The last few years have seen an increased number of metabolic pathway design and engineering in P. pastoris, mainly towards the production of complex (secondary) metabolites. In this review, we discuss the potential role of P. pastoris as a platform for metabolic engineering, its strengths, and major requirements for future developments of chassis strains based on synthetic biology principles.

Orthologous promoters from related methylotrophic yeasts surpass expression of endogenous promoters of Pichia pastoris

“…Over the past three decades, the methylotrophic yeasts Pichia pastoris (Pp), Hansenula polymorpha (Hp), Candida boidinii (Cb) and Pichia methanolica (Pm) have emerged as powerful alternatives, enabling high cell density fermentation and simple, pure secretion of heterologous proteins (Gellissen 2000;Hartner and Glieder 2006;Yurimoto et al 2011;). The two Pichia species and H. polymorpha have phylogenetically been reassigned as Komagataella and Ogataea species, respectively resulting in the formal names Komagataella phaffii, Ogataea polymorpha, and Ogataea methanolica (Peña et al 2018). Amongst these four methylotrophic yeasts, P. pastoris is most commonly applied for heterologous protein production, even surpassing S. cerevisiae according to a recent literature survey (Bill 2014).…”

Engineering substrate and energy metabolism for living cell production of cytidine‐5′‐diphosphocholine

“…It is a widely used industrial microorganism and has several other advantages over other eukaryotic and prokaryotic expression hosts, such as simple medium, easy genetic manipulation, diverse posttranslational modifications, less extensive glycosylation, good secretion ability (Safder et al, 2018). Till date, a variety of proteins, small molecule drugs, and food additives have been synthesized using P. pastoris as the chassis (Peña et al, 2018;Safder et al, 2018).…”

Pichia pastoris as a Versatile Cell Factory for the Production of Industrial Enzymes and Chemicals: Current Status and Future Perspectives

“…In contrast to its well‐known function as a protein expression system, the potential of P. pastoris as a cell factory for the production of chemicals has been largely underestimated. Nevertheless, increasing number of studies reported using P. pastoris as a microbial cell factory for the production of chemicals either via biocatalysis or more recently via fermentation . This paper aims to outline the major progress made in recent years toward developing P. pastoris as an efficient yeast cell factory with emphasis on its role as a whole‐cell catalyst for chemical production.…”